mouse anti extracellular human tshr antibody  (Bio-Rad)

Journal: The FASEB Journal

Article Title: Signaling-sensitive amino acids surround the allosteric ligand binding site of the thyrotropin receptor

doi: 10.1096/fj.09-149146

Figure Lengend Snippet: Orthosteric and allosteric binding sites of the TSHR. TSHR is characterized by a unique structural feature compared with other family A GPCRs: a large extracellular region (∼400 aa), which can be subdivided into two parts: the LRRD (light orange) and the cysteine-rich hinge region (indicated by dotted line because no structure or homology model is available). Together, both extracellular parts are responsible for hormone binding and hormone selectivity. The hormone TSH consists of an α-subunit (cyan) and a β-subunit (lilac). In contrast, the small synthetic ligand (LMW-Ag, blue) is shown to bind allosterically in a pocket (yellow surface) located in the heptahelical transmembrane region (orange backbone) of the TSHR. The pocket is characterized by specific biophysical properties complementary to that of the ligand. Simultaneous binding of both ligands to the same receptor has not been shown to occur.

Article Snippet: Then they were incubated for 30 min with a 1:200 dilution of a mouse anti-extracellular human TSHR antibody (2C11; Serotec, Oxford, UK) in FACS buffer.

Techniques: Binding Assay

Journal: The FASEB Journal

Article Title: Signaling-sensitive amino acids surround the allosteric ligand binding site of the thyrotropin receptor

doi: 10.1096/fj.09-149146

Figure Lengend Snippet: Newly identified CAMs in the thyrotropin receptor surround the allosteric binding site of a LMW agonist. Left panel: molecular homology model of the serpentine domain (aa C408–I698) without the large N-terminal tail of the human TSHR is based on the crystal structure of inactive bovine rhodopsin. Backbone (white) of the TSHR and the previously identified CAMs (magenta) I640V (TMH6) and I568V (ECL2) are displayed. Nine positions of the newly identified CAMs (red sticks) are localized in the upper part of the TMHs toward the extracellular loops. Right panel: closeup view of the activated TSHR conformation, which is based on the opsin crystal structure, occupied by LMW-Ag (11). Wild-type amino acids of the 9 CAM positions (red) are arranged in a cluster. They are spatially close or interact directly with each other and surround the allosteric ligand binding pocket of LMW-Ag (blue). Therefore, these amino acids may be involved in TSHR activation by small-molecule ligands.

Article Snippet: Then they were incubated for 30 min with a 1:200 dilution of a mouse anti-extracellular human TSHR antibody (2C11; Serotec, Oxford, UK) in FACS buffer.

Techniques: Binding Assay, Ligand Binding Assay, Activation Assay